Dynamic Behavior of FCHO1 Revealed by Live-Cell Imaging Microscopy: Its Possible Involvement in Clathrin-Coated Vesicle Formation

Sakaushi, Shinji; Inoue, Koichiro; Zushi, Hitomi; Senda-Murata, Kaori; Fukada, Takashi; Oka, Saori; Sugimoto, Kenji

doi:10.1271/bbb.60720

Cited by 20 publications

(12 citation statements)

References 12 publications

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“…The μ-homology region in the Syp1/FCHO1 family is related to the μ chain-C-terminal segment that binds to tyrosine sorting motifs found in membrane cargo collected in clathrin coated pits [49-51]. Similar to Syp1, FCHO1 is also associated with clathrin-coated pits in animal cells [52]. The early arrival of Syp1 at the endocytic patch, diminished localization in the absence of clathrin, and presence of a μ-homology region, suggests that Syp1 could be a cargo specific adaptor [36].…”

Section: Discussionmentioning

confidence: 99%

The F-BAR Protein Syp1 Negatively Regulates WASp-Arp2/3 Complex Activity during Endocytic Patch Formation

et al. 2009

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Background Actin polymerization by Arp2/3 complex must be tightly regulated to promote endocytosis. While many Arp2/3 complex activators have been identified, mechanisms for its negative regulation have remained more elusive. We analyzed the yeast arp2-7 allele, which is biochemically unique in causing unregulated actin assembly in vitro in the absence of Arp2/3 activators. Results We examined endocytosis in arp2-7 mutants by live cell imaging of Sla1-GFP, a coat marker, and Abp1-RFP, which marks the later actin phase of endocytosis. Sla1-GFP and Abp1-RFP lifetimes were accelerated in arp2-7 mutants, which is opposite to actin nucleation-impaired arp2 alleles or deletions of Arp2/3 activators. We performed a screen for multi-copy suppressors of arp2-7 and identified SYP1, a FCHO1 homologue, which contains F-BAR and AP-2μ homology domains. Over-expression of SYP1 in arp2-7 cells slowed Sla1-GFP lifetimes closer to wild type cells. Further, purified Syp1 directly inhibited Las17/WASp stimulation of Arp2/3 complex-mediated actin assembly. This activity was mapped to a fragment of Syp1 located between its F-BAR and AP-2μ homology domains, and depends on non-VCA sequences in Las17/WASp. Conclusions Together, these data identify Syp1 as a novel negative regulator of WASp-Arp2/3 complex that helps choreograph the precise timing of actin assembly during endocytosis.

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Section: Discussionmentioning

confidence: 99%

The F-BAR Protein Syp1 Negatively Regulates WASp-Arp2/3 Complex Activity during Endocytic Patch Formation

et al. 2009

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“…This hypothesis in particular addressed the earliest stages of coat invagination and it was proposed that FCHo, a curvature-inducing F-BAR protein (Henne et al 2007;Shimada et al 2007) implicated in clathrin-mediated endocytosis (Sakaushi et al 2007), might induce a "pimple" in the plasma membrane and so initiate the formation of a curved clathrin coat (Henne et al 2010). However, subsequent experiments in zebrafish embryos and mammalian cultured cells suggested FCHo is nonessential for general CME (Shimada et al 2007;Cocucci et al 2012) and is rather an adaptor for BMP receptor (Umasankar et al 2012) and/or facilitates CCP invagination Shimada et al 2007;Cocucci et al 2012).…”

Section: Eaps and Endocytic Site Nucleation: Whence The Pits?mentioning

confidence: 99%

Endocytic Accessory Factors and Regulation of Clathrin-Mediated Endocytosis

Merrifield

Kaksonen

2014

Cold Spring Harbor Perspectives in Biology

118

111

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“…Similarity searching with the Syp1p sequence produced hits against a number of conserved metazoan proteins, including the endocytic protein SGIP1-␣, the putative endocytic F-BAR protein FCHo1, and the F-BAR protein FCHo2 (Figure 2A; Henne et al, 2007;Sakaushi et al, 2007;Uezu et al, 2007).…”

Section: Syp1p Is a Conserved Endocytic Proteinmentioning

confidence: 99%

Early-Arriving Syp1p and Ede1p Function in Endocytic Site Placement and Formation in Budding Yeast

Stimpson

Toret

Cheng

et al. 2009

MBoC

122

151

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Recent studies have revealed the detailed timing of protein recruitment to endocytic sites in budding yeast. However, little is understood about the early stages of their formation. Here we identify the septin-associated protein Syp1p as a component of the machinery that drives clathrin-mediated endocytosis in budding yeast. Syp1p arrives at endocytic sites early in their formation and shares unique dynamics with the EH-domain protein Ede1p. We find that Syp1p is related in amino acid sequence to several mammalian proteins one of which, SGIP1-␣, is an endocytic component that binds the Ede1p homolog Eps15. Like Syp1p, SGIP1-␣ arrives early at sites of clathrin-mediated endocytosis, suggesting that Syp1p/Ede1p and SGIP1-␣/Eps15 may have a conserved function. In yeast, both Syp1p and Ede1p play important roles in the rate of endocytic site turnover. Additionally, Ede1p is important for endocytic site formation, whereas Syp1p acts as a polarized factor that recruits both Ede1p and endocytic sites to the necks of emerging buds. Thus Ede1p and Syp1p are conserved, early-arriving endocytic proteins with roles in the formation and placement of endocytic sites, respectively. INTRODUCTIONThe dynamics of protein recruitment to sites of clathrinmediated endocytosis have been revealed by live-cell microscopy in budding yeast and mammalian cells (Merrifield et al., 2002;Kaksonen et al., 2003;Kaksonen et al., 2005). These studies have identified numerous proteins that sequentially assemble at endocytic sites and have shown that actin polymerization can power internalization. It is now evident that the dynamic recruitment and disappearance of endocytic proteins are precisely coordinated for productive internalization and that each protein has defined dynamics at endocytic sites. In Saccharomyces cerevisiae four endocytic modules have been defined that each contain proteins with similar dynamics: the coat, WASP/myo, amphiphysin, and actin modules (Kaksonen et al., 2005).Despite detailed knowledge of events at endocytic sites, little is understood about the early stages of their formation. The best candidates for proteins that initiate endocytic site formation are those that arrive earliest. In mammalian cells the classical coat protein clathrin marks the earliest known stage of endocytic site formation (Merrifield et al., 2002). Additionally, the adapter AP-2, is critical for site formation and has similar dynamic behavior to clathrin and so is thought to arrive early (Hinrichsen et al., 2003;Motley et al., 2003;Ehrlich et al., 2004;Keyel et al., 2004). In yeast, the role of AP-2 is unclear, but clathrin, a coat module component, marks the first stages of endocytosis, and its deletion causes severe defects in the number of sites formed (Huang et al., 1999;Kaksonen et al., 2005;Newpher et al., 2005;Newpher and Lemmon, 2006). An additional yeast protein, Ede1p, arrives early and plays a role in endocytic site formation, although its dynamics and function have yet to be fully investigated (Kaksonen et al., 2005;Toshima et al., 200...

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Dynamic Behavior of FCHO1 Revealed by Live-Cell Imaging Microscopy: Its Possible Involvement in Clathrin-Coated Vesicle Formation

Cited by 20 publications

References 12 publications

The F-BAR Protein Syp1 Negatively Regulates WASp-Arp2/3 Complex Activity during Endocytic Patch Formation

The F-BAR Protein Syp1 Negatively Regulates WASp-Arp2/3 Complex Activity during Endocytic Patch Formation

Endocytic Accessory Factors and Regulation of Clathrin-Mediated Endocytosis

Early-Arriving Syp1p and Ede1p Function in Endocytic Site Placement and Formation in Budding Yeast

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